Antigen retrieval reagent

ImmunoSaver

ImmunoSaver is an antigen retrieval reagent for formalin-fixed tissues. This product is used to restore antigenicity that was reduced by formalin fixation. As a result, the quality of immunostaining is improved. While various conditions have to be examined in conventional methods, this product can retrieve a wide range of antigens under the same conditions.

Features
Composition
Usage
Example of staining
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Features

It restores the antigenicity of formalin-fixed tissues.
It improves the quality of immunostaining.
It retrieves a wide range of antigens under the same conditions.

Composition

10% sodium citraconate solution

Usage

Dilute ImmunoSaver by a factor of 200 with distilled or deionized water, and heat to 98°C.
Place deparaffinized tissue sections in the diluted solution, and incubate at 98°C for 45 minutes.
Wash the tissue sections with PBS three times for 5 minutes each, and proceed to immunostaining.

Example of staining

Treat formalin-fixed human pancreatic paraffin sections with ImmunoSaver.
Wash the paraffin sections once with PBST (PBS containing Triton X-100 at a concentration of 0.2%) and twice with PBS.
Block at room temperature for 1 hour.
Rinse the sections gently with PBS and react them with the primary antibody diluted in PBST at 4°C overnight. (rabbit anti SNAP-25 1/8,000)
Wash the sections once with PBST and twice with PBS.
React the sections with the biotin-labeled secondary antibody at room temperature for 1 hour.
Wash the sections once with PBST and twice with PBS.
React the sections with peroxidase-labeled avidin (1/400, diluted in PBS) at room temperature for 1 hour.
Wash the sections once with PBST and twice with PBS.
Use DAB as substrate for chromogenic reaction.
Wash the sections with distilled water for 5 minutes, three times.
Stain the sections with Mayer's acidic hemalum solution for approximately 1 to 3 minutes.
Wash the sections with running water, rinse with distilled water two or three times, and discriminate color with 70% ethanol.
Clear and mount the sections.

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For research use or further manufacturing use only. Not for use in diagnostic procedures.

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